Abstract:
:Improvements in proteome analysis of erythrocyte membrane proteins by two-dimensional electrophoresis are here reported. In particular, a differential extraction procedure was set up allowing separation of integral membrane proteins from peripheral species. Moreover, the use of dilute Immobiline gels (down to as low as 3% T matrix) permitted a better penetration and transfer inside the gel of proteins with large M(r). These protocol modifications, combined with sample delipidation and alkylation prior to electrophoresis, which prevented generation of homo- and hetero-oligomers following disulfide scrambling phenomena, allowed the display of more than 500 spots in the pI/M(r) plane. Among those, noteworthy was the presence of high levels of filamentous proteins, such as alpha-spectrin and ankyrins, or integral membrane proteins, such as band 3, band 4.1 and 4.2, not displayed or barely present in other maps exploiting immobilized pH gradients in the first dimension. Accordingly, our results show that this 2D mapping technique is a valuable tool in exploring pathologies related to genetic defects associated to membrane proteins.
journal_name
J Proteome Resjournal_title
Journal of proteome researchauthors
Bruschi M,Seppi C,Arena S,Musante L,Santucci L,Balduini C,Scaloni A,Lanciotti M,Righetti PG,Candiano Gdoi
10.1021/pr050026vkeywords:
subject
Has Abstractpub_date
2005-07-01 00:00:00pages
1304-9issue
4eissn
1535-3893issn
1535-3907journal_volume
4pub_type
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